Brief statement of overall objectives of the total project: A DNA-directed cell-free system for RNA and protein synthesis is being used to quantitatively analyze the factors necessary for regulation of genes associated with the lac and ara operons. The factors which are being studied in this synthetic system or in more purified systems include the catabolite gene activator protein, CAP, the araC protein and magic spot, ppGpp. The characterization of a small RNA molecule of unknown function whose synthesis is strongly inhibited by magic spot is also being investigated. The discovery of this RNA molecule came as a surprise; it is suspected that it may be involved in the regulation of DNA synthesis.